US4889818A - Purified thermostable enzyme - Google Patents
Purified thermostable enzyme Download PDFInfo
- Publication number
- US4889818A US4889818A US07/063,509 US6350987A US4889818A US 4889818 A US4889818 A US 4889818A US 6350987 A US6350987 A US 6350987A US 4889818 A US4889818 A US 4889818A
- Authority
- US
- United States
- Prior art keywords
- dna
- sequence
- enzyme
- nucleic acid
- polymerase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
- C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1276—RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
Abstract
Description
Primer 1: 3'GGGGGGGGGG 5'
Primer 2: 5'AAAAAAAAAA 3'
______________________________________ Number of Double Strands After 0 to n Cycles Long Specific Cycle Number Template Products Sequence [S] ______________________________________ 0 1 -- -- 1 1 1 0 2 1 2 1 3 1 3 4 5 1 5 26 10 1 10 1013 15 1 15 32,752 20 1 20 1,048,555 n 1 n (2.sup.n -n-1) ______________________________________
5'-ACACAACTGTGTTCACTAGC-3' (PC03)
5'-CAACTTCATCCACGTTCACC-3' (PC04)
______________________________________ Sodium Citrate 1 mM Potassium Phosphate, pH 7.9 5 mM Ammonium Chloride 10 mM Magnesium Sulfate 0.2 mM Calcium Chloride 0.1 mM Sodium Chloride 1 g/l Yeast Extract 1 g/l Tryptone 1 g/l Glucose 2 g/l Ferrous Sulfate 0.01 mM ______________________________________
5'-*CCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG-3' (RS24)
5'-*CTCCTGAGGAGAAGTCTGC-3' (RS18)
5'-CTTCTGcagCAACTGTGTTCACTAGC-3' (GH18)
5'-CACaAgCTTCATCCACGTTCACC-3' (GH19)
TABLE 1 ______________________________________ Expression of a Thermostable DNA Polymerase Activity in E. coli# Units*/ml Sample IPTG +IPTG ______________________________________ BSM13/DG98- -- 0.02 pFC82/DG98 2.2 2.7 pFC85/DG98 11.9 643.8 ______________________________________ #Cells were grown to late log phase (+/-IPTG, 10 mM), harvested, sonicated, heated at 75° C. for 20 minutes, centrifuged and the clarified supernatant assayed at 70° C. for DNA polymerase activity. *1 unit = 1 nM dCTP incorporated in 30 minutes.
5'-CATGCCTCTTTGCACCATTC-3'(RS79) and
5'-TGGTAGCTGGATTGTAGCTG-3'(RS80)
5'-ATAATCCACCTATCCCAGTAGGAGAAAT-3'(SK38) and
5'-TTTGGTCCTTGTCTTATGTCCAGAATGC-3'(SK39)
______________________________________ Deposit Designation CMCC No. ATCC No. Deposit ______________________________________ CH35:Taq #4-2 3125 ATCC 40336 5/28/87 E. coli DG98/ 3128 ATCC 67422 5/28/87 pFC83 E. coli DG98/ 3127 ATCC 67421 5/28/87 pFC85 ______________________________________
Claims (3)
Priority Applications (54)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US07/063,509 US4889818A (en) | 1986-08-22 | 1987-06-17 | Purified thermostable enzyme |
CA000608796A CA1340921C (en) | 1986-08-22 | 1987-07-17 | Purified thermostable enzyme |
CA000542406A CA1338457C (en) | 1986-08-22 | 1987-07-17 | Purified thermostable enzyme |
NZ221517A NZ221517A (en) | 1986-08-22 | 1987-08-20 | Thermostable enzyme: thermus aquaticus dna polymerase, and processes for amplifying, detecting and/or cloning nucleic acid sequences using said enzyme |
NO873546A NO305488B1 (en) | 1986-08-22 | 1987-08-21 | Thermostable DNA polymerase, DNA sequence encoding it, vector comprising the DNA sequence, microorganism transformed with the DNA sequence and preparations containing the thermostable DNA polymerase |
BR8704332A BR8704332A (en) | 1986-08-22 | 1987-08-21 | PURIFIED THERMOSTABLE ENZYME, ENZYME GENE CODING, SELECTED PLASMIDIOUS, BACTEREOPHOG, STABLE ENZYME COMPOSITION, PROCESS TO AMPLIFY AT LEAST ONE SPECIFIC SEQUENCE OF NUCLEIC ACID, PROCESS OF A DETENTION OF A DETECTIVE DECAY DETECT PRESENCE OR ABSENCE OF AT LEAST ONE VARIATION OF NUCLEOTIDEOS IN SEQUENCE IN ONE OR MORE NUCLEIC ACIDS, PROCESS FOR CLONING IN CLONING VECTOR OF ONE OR MORE SPECIFIC NUCLEIC ACID AND AMPLIFIED NUCLEIC ACID SEQUENCES |
AT96119077T ATE391771T1 (en) | 1986-08-22 | 1987-08-21 | STABLE ENZYME COMPOSITION CONTAINING A THERMOSTABLE NUCLEIC ACID POLYMERASE |
ES96119077T ES2105998T1 (en) | 1986-08-22 | 1987-08-21 | STABLE ENZYMATIC COMPOSITION THAT INCLUDES A THERMOSTABLE NUCLEIC ACID POLYMERASE ENZYME. |
DE3752073T DE3752073T3 (en) | 1986-08-22 | 1987-08-21 | Purified thermostable enzyme and method for amplifying, detecting and / or cloning nucleic acid sequences using this enzyme |
CN198787105787A CN87105787A (en) | 1986-08-22 | 1987-08-21 | The thermostability enzyme of purifying and with its increase, the method for detection and/or cloning nucleic acid sequences |
EP96119077.4A EP0776970B2 (en) | 1986-08-22 | 1987-08-21 | Stable enzyme composition comprising a thermostable nucleic acid polymerase enzyme |
IE19970680A IE970680A1 (en) | 1986-08-22 | 1987-08-21 | Stable enzyme composition comprising a thermostable nucleic acid polymerase enzyme |
AU77298/87A AU7729887A (en) | 1986-08-22 | 1987-08-21 | Purified thermostable enzyme |
ES87307433T ES2104550T5 (en) | 1986-08-22 | 1987-08-21 | THERMOSTABLE PURIFIED ENZYME AND PROCESS TO EXPAND, DETECT AND / OR CLONE SEQUENCES OF NUCLEIC ACID USING THIS ENZYME. |
AT87307433T ATE154072T1 (en) | 1986-08-22 | 1987-08-21 | PURIFIED THERMOSTABLE ENZYME AND METHOD FOR AMPLIFICATION, DETECTION AND/OR CLONING NUCLEIC ACID SEQUENCES USING SUCH ENZYME |
DE0776970T DE776970T1 (en) | 1986-08-22 | 1987-08-21 | Stable enzyme composition containing a thermostable nucleic acid polymerase |
IL10303987A IL103039A (en) | 1986-08-22 | 1987-08-21 | Process for amplifying, detecting and/or cloning of specific nucleic acid sequences using a thermostable enzyme |
DE198787307433T DE258017T1 (en) | 1986-08-22 | 1987-08-21 | CLEANED THERMOSTABLE ENZYME AND METHOD FOR REINFORCING, DETECTING AND / OR CLONING NUCLEIC ACID SEQUENCES BY USING THIS ENZYME. |
EP87307433A EP0258017B2 (en) | 1986-08-22 | 1987-08-21 | Purified thermostable enzyme and process for amplifying, detecting, and/or cloning nucleic acid sequences using said enzyme |
IL83605A IL83605A0 (en) | 1986-08-22 | 1987-08-21 | Purified thermostable enzyme and process for amplifying,detecting,and/or cloning nucleic acid sequences using said enzyme |
DK198704384A DK175806B1 (en) | 1986-08-22 | 1987-08-21 | Purified thermostable enzyme and method for amplification, detection and / or cloning of the nucleic acid sequence using this enzyme |
SG1996007005A SG46644A1 (en) | 1986-08-22 | 1987-08-21 | Purified thermostable enzyme and process for amplifying detecting and/or cloning nucleic acid sequences using said enzyme |
DE3752392T DE3752392T3 (en) | 1986-08-22 | 1987-08-21 | Stable enzyme composition containing a thermostable nucleic acid polymerase |
KR1019870009197A KR960016559B1 (en) | 1986-08-22 | 1987-08-22 | Thermostable enzyme, thermus aquaticus dna polymerase, and processes for amplifying, detecting |
FI875543A FI875543A (en) | 1987-06-17 | 1987-12-16 | RENAT THERMOSTABILT ENZYM OCH FOERFARANDE FOER AMPLIFIERING, DETEKTERING OCH / ELLER KLONING AV NUCLEIN SYRASE SEQUENTOR GENOM ANVAENDNING AV DETTA ENZYM. |
HU608987A HUT45555A (en) | 1987-06-17 | 1987-12-29 | Purified, heat-resisting enzyme and process for strengthening, detecting and cloning sequences of nukleinic acides utilizing former enzyme |
JP1003887A JPH0824570B2 (en) | 1986-08-22 | 1989-01-12 | Gene encoding thermostable DNA polymerase |
CA000608780A CA1333891C (en) | 1986-08-22 | 1989-08-18 | Purified thermostable enzyme |
CA000608795A CA1337405C (en) | 1986-08-22 | 1989-08-18 | Purified thermostable enzyme |
US07/523,394 US5079352A (en) | 1986-08-22 | 1990-05-15 | Purified thermostable enzyme |
US07/567,244 US5374553A (en) | 1986-08-22 | 1990-08-13 | DNA encoding a thermostable nucleic acid polymerase enzyme from thermotoga maritima |
AU63910/90A AU632857C (en) | 1986-08-22 | 1990-10-08 | Purified thermostable enzyme |
JP2326114A JP2719529B2 (en) | 1986-08-22 | 1990-11-29 | Thermostable nucleic acid polymerase composition |
JP2326113A JPH03180178A (en) | 1986-08-22 | 1990-11-29 | Heat stabilized dna polymerase derivative |
US07/977,434 US5466591A (en) | 1986-08-22 | 1991-09-30 | 5' to 3' exonuclease mutations of thermostable DNA polymerases |
US07/873,897 US6127155A (en) | 1986-08-22 | 1992-04-24 | Stabilized thermostable nucleic acid polymerase compositions containing non-ionic polymeric detergents |
IL103039A IL103039A0 (en) | 1986-08-22 | 1992-09-03 | Process for amplifying,detecting and/or cloning of specific nucleic acid sequences using a thermostable enzyme |
US07/971,798 US5352600A (en) | 1986-08-22 | 1992-11-05 | Purified thermostable enzyme |
NO930220A NO301890B1 (en) | 1986-08-22 | 1993-01-22 | Method for amplifying at least one specific nucleic acid sequence and using the method |
US08/080,243 US5407800A (en) | 1986-08-22 | 1993-06-17 | Reverse transcription with Thermus thermophilus polymerase |
US08/082,182 US5310652A (en) | 1986-08-22 | 1993-06-24 | Reverse transcription with thermostable DNA polymerase-high temperature reverse transcription |
US08/113,531 US5455170A (en) | 1986-08-22 | 1993-08-27 | Mutated thermostable nucleic acid polymerase enzyme from Thermus species Z05 |
US08/119,754 US5405774A (en) | 1986-08-22 | 1993-09-10 | DNA encoding a mutated thermostable nucleic acid polymerase enzyme from thermus species sps17 |
JP6002160A JP2502042B2 (en) | 1986-08-22 | 1994-01-13 | Method for producing thermostable DNA polymerase |
JP6002142A JP2502041B2 (en) | 1986-08-22 | 1994-01-13 | Nucleic acid amplification method using thermostable DNA polymerase |
US08/199,509 US5618703A (en) | 1986-08-22 | 1994-02-22 | Unconventional nucleotide substitution in temperature selective RT-PCR |
US08/311,612 US5641864A (en) | 1986-08-22 | 1994-09-22 | Kits for high temperature reverse transcription of RNA |
US08/384,817 US5693517A (en) | 1987-06-17 | 1995-02-02 | Reagents and methods for coupled high temperature reverse transcription and polymerase chain reactions |
US08/384,490 US5618711A (en) | 1986-08-22 | 1995-02-06 | Recombinant expression vectors and purification methods for Thermus thermophilus DNA polymerase |
US08/449,050 US5561058A (en) | 1986-08-22 | 1995-05-24 | Methods for coupled high temperatures reverse transcription and polymerase chain reactions |
US08/459,383 US5789224A (en) | 1986-08-22 | 1995-06-02 | Recombinant expression vectors and purification methods for thermus thermophilus DNA polymerase |
US08/475,231 US5624833A (en) | 1986-08-22 | 1995-06-07 | Purified thermostable nucleic acid polymerase enzyme from Thermotoga maritima |
US08/520,422 US5674738A (en) | 1986-08-22 | 1995-08-29 | DNA encoding thermostable nucleic acid polymerase enzyme from thermus species Z05 |
GR970402257T GR3024616T3 (en) | 1986-08-22 | 1997-09-03 | Purified thermostable enzyme and process for amplifying, detecting, and/or cloning nucleic acid sequences using said enzyme |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US89924186A | 1986-08-22 | 1986-08-22 | |
US07/063,509 US4889818A (en) | 1986-08-22 | 1987-06-17 | Purified thermostable enzyme |
Related Parent Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US89924186A Continuation-In-Part | 1986-08-22 | 1986-08-22 | |
US89924186A Continuation | 1986-08-22 | 1986-08-22 | |
US08899241 Continuation-In-Part | 1996-08-22 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14344188A Continuation-In-Part | 1986-08-22 | 1988-01-12 | |
US14344188A Continuation | 1986-08-22 | 1988-01-12 |
Publications (1)
Publication Number | Publication Date |
---|---|
US4889818A true US4889818A (en) | 1989-12-26 |
Family
ID=26743490
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US07/063,509 Expired - Lifetime US4889818A (en) | 1986-08-22 | 1987-06-17 | Purified thermostable enzyme |
Country Status (1)
Country | Link |
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US (1) | US4889818A (en) |
Cited By (628)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4965188A (en) * | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
WO1991002090A1 (en) * | 1989-08-03 | 1991-02-21 | Promega Corporation | Modified taq dna polymerase |
EP0455430A2 (en) | 1990-04-26 | 1991-11-06 | New England Biolabs, Inc. | Purified thermostable DNA polymerase obtainable from Thermococcus litoralis |
WO1991017239A1 (en) * | 1990-05-03 | 1991-11-14 | Cornell Research Foundation, Inc. | A thermostable ligase mediated dna amplification system for the detection of genetic diseases |
US5066584A (en) * | 1988-09-23 | 1991-11-19 | Cetus Corporation | Methods for generating single stranded dna by the polymerase chain reaction |
US5079352A (en) * | 1986-08-22 | 1992-01-07 | Cetus Corporation | Purified thermostable enzyme |
WO1992009689A1 (en) * | 1990-12-03 | 1992-06-11 | Stratagene | PURIFIED THERMOSTABLE $i(PYROCOCCUS FURIOSUS) |
US5122345A (en) * | 1988-07-12 | 1992-06-16 | President And Fellows Of Harvard College | Dna sequencing apparatus |
EP0519338A1 (en) | 1991-06-20 | 1992-12-23 | F. Hoffmann-La Roche Ag | Improved methods for nucleic acid amplification |
US5192674A (en) * | 1988-08-26 | 1993-03-09 | Toyo Boseki Kabushiki Kaisha | Thermostable dna polymerase thermus thermophilus and a method of producing the same |
US5196305A (en) * | 1989-09-12 | 1993-03-23 | Eastman Kodak Company | Diagnostic and amplification methods using primers having thymine at 3' end to overcome primer-target mismatch at the 3' end |
US5210036A (en) * | 1990-04-26 | 1993-05-11 | New England Biolabs, Inc. | Purified thermostable DNA polymerase obtainable from thermococcus litoralis |
US5210015A (en) * | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
US5219727A (en) * | 1989-08-21 | 1993-06-15 | Hoffmann-Laroche Inc. | Quantitation of nucleic acids using the polymerase chain reaction |
US5242818A (en) * | 1988-08-26 | 1993-09-07 | Toyo Boseki Kabushiki Kaisha | Method of producing a thermostable DNA polymerase from Thermus thermophilus |
US5268289A (en) * | 1991-12-27 | 1993-12-07 | Epicentre Technologies Corp. | Thermostable ribonuclease H |
US5270179A (en) * | 1989-08-10 | 1993-12-14 | Life Technologies, Inc. | Cloning and expression of T5 DNA polymerase reduced in 3'- to-5' exonuclease activity |
US5308751A (en) * | 1992-03-23 | 1994-05-03 | General Atomics | Method for sequencing double-stranded DNA |
US5310893A (en) * | 1986-03-31 | 1994-05-10 | Hoffmann-La Roche Inc. | Method for HLA DP typing |
US5310652A (en) * | 1986-08-22 | 1994-05-10 | Hoffman-La Roche Inc. | Reverse transcription with thermostable DNA polymerase-high temperature reverse transcription |
US5316082A (en) * | 1992-08-28 | 1994-05-31 | Mobil Oil Corporation | Method of effectively diverting treating fluid from a high permeability interval during well stimulation |
US5322770A (en) * | 1989-12-22 | 1994-06-21 | Hoffman-Laroche Inc. | Reverse transcription with thermostable DNA polymerases - high temperature reverse transcription |
US5333675A (en) * | 1986-02-25 | 1994-08-02 | Hoffmann-La Roche Inc. | Apparatus and method for performing automated amplification of nucleic acid sequences and assays using heating and cooling steps |
US5338671A (en) * | 1992-10-07 | 1994-08-16 | Eastman Kodak Company | DNA amplification with thermostable DNA polymerase and polymerase inhibiting antibody |
US5352600A (en) * | 1986-08-22 | 1994-10-04 | Hoffmann-La Roche Inc. | Purified thermostable enzyme |
US5352778A (en) * | 1990-04-26 | 1994-10-04 | New England Biolabs, Inc. | Recombinant thermostable DNA polymerase from archaebacteria |
US5374553A (en) * | 1986-08-22 | 1994-12-20 | Hoffmann-La Roche Inc. | DNA encoding a thermostable nucleic acid polymerase enzyme from thermotoga maritima |
US5403707A (en) * | 1993-05-14 | 1995-04-04 | Eastman Kodak Company | Diagnostic compositions, elements, methods and test kits for amplification and detection of retroviral DNA using primers having matched melting temperatures |
US5405774A (en) * | 1986-08-22 | 1995-04-11 | Hoffmann-La Roche Inc. | DNA encoding a mutated thermostable nucleic acid polymerase enzyme from thermus species sps17 |
US5407800A (en) * | 1986-08-22 | 1995-04-18 | Hoffmann-La Roche Inc. | Reverse transcription with Thermus thermophilus polymerase |
US5411886A (en) * | 1990-10-29 | 1995-05-02 | Nihon Shokuhin Kako Co., Ltd. | Xylose isomerase gene of Thermus aquaticus |
US5411876A (en) * | 1990-02-16 | 1995-05-02 | Hoffmann-La Roche Inc. | Use of grease or wax in the polymerase chain reaction |
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US5455170A (en) * | 1986-08-22 | 1995-10-03 | Hoffmann-La Roche Inc. | Mutated thermostable nucleic acid polymerase enzyme from Thermus species Z05 |
US5459055A (en) * | 1991-12-27 | 1995-10-17 | Epicentre Technologies Corporation | Thermostable ribonuclease H isolated from Thermus flavus |
US5466591A (en) * | 1986-08-22 | 1995-11-14 | Hoffmann-La Roche Inc. | 5' to 3' exonuclease mutations of thermostable DNA polymerases |
US5474920A (en) * | 1993-11-23 | 1995-12-12 | State Of Oregon, Acting By And Through The Oregon State Board Of Higher Education On Behalf Of The Oregon Health Sciences University | Modified thermo-resistant DNA polymerases |
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